Department of Biological Sciences, University of Essex, Colchester, Essex CO4 3SQ, UK.
Anal Biochem. 2011 May 15;412(2):183-8. doi: 10.1016/j.ab.2011.01.036. Epub 2011 Feb 1.
Chromatin immunoprecipitation (ChIP) is an important technique in the study of DNA/protein interactions. The ChIP procedure, however, has limitations in that it is lengthy, can be inconsistent, and is prone to nonspecific binding of DNA and proteins to the bead-based solid-phase matrices that are often used for the immunoprecipitation step. In this investigation, we examined the utility of a new matrix for ChIP assays, BioVyon Protein A, a solid support based on porous polyethylene. In ChIP experiments carried out using two antibodies and seven DNA loci, the performance of BioVyon Protein A was significantly better, with a greater percentage of DNA pull-down in all of the assays tested compared with bead-based matrices, Protein A Sepharose, and Dynabeads Protein A. Furthermore, the rigid porous disc format within a column made the BioVyon matrix much easier to use with fewer steps and less equipment requirements, resulting in a significant reduction in the time taken to process the ChIP samples. In summary, BioVyon Protein A provides a column-based assay method for ChIP and other immunoprecipitation-based procedures; the rigid porous structure of BioVyon enables a fast and robust protocol with higher ChIP enrichment ratios.
染色质免疫沉淀（ChIP）是研究 DNA/蛋白质相互作用的重要技术。然而，ChIP 程序存在一些局限性，例如过程冗长、结果不一致，并且在免疫沉淀步骤中，DNA 和蛋白质经常会非特异性地结合到基于珠子的固相基质上。在这项研究中，我们研究了一种新型 ChIP 检测试剂盒，BioVyon Protein A 的应用，这是一种基于多孔聚乙烯的固相载体。在使用两种抗体和七个 DNA 基因座进行的 ChIP 实验中，BioVyon Protein A 的性能明显更好，与基于珠子的基质、Protein A Sepharose 和 Dynabeads Protein A 相比，所有检测的 DNA 下拉百分比都更高。此外，柱内的刚性多孔盘结构使得 BioVyon 基质的使用更加简便，所需步骤和设备更少，从而显著减少了 ChIP 样品处理所需的时间。总之，BioVyon Protein A 为 ChIP 及其它免疫沉淀法提供了一种基于柱的检测方法；BioVyon 的刚性多孔结构可实现快速而稳健的方案，具有更高的 ChIP 富集率。
